LC-API/MS in Drug Metabolism and Pharmacokinetic Studies

Imrie, G A (2007) LC-API/MS in Drug Metabolism and Pharmacokinetic Studies. Doctoral thesis, University of Sunderland.

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The use of API interfaces with quadrupole mass spectrometers has been shown to give rise to good sensitivity, selectivity, and robustness for the interfacing of LC to MS. Since their introduction in the 1990s the technique has rapidly become widespread, but at the outset of this research programme, there were still a number of problems associated with it, particularly when dealing with complex sample matrices. The aim of this research programme was to study illustrative examples of the kinds of problems associated with the analysis of biological samples using LC-API-MS in an attempt to arrive at strategies which could be employed to eliminate, or at least compensate for, the problems.

Commonly reported problems include the occurrence of matrix effects - a change in response of the target analyte(s) as a result of the presence in the samples of co /late eluting interferences. An investigation which compared ESI with APCI ionisation illustrated a significant drawback in the accepted methodology for the elimination of matrix effects.

Optimal LC conditions for a number of assays may use non-MS-friendly mobile phases. A simple and convenient solution to this problem was found to be the post column addition of organic modifier, which reproducibly and reliably enhanced sensitivity. This approach was initially used for a range of dihydropyridine calcium channel blockers and was subsequently applied to a range of chiral compounds from different therapeutic groups to illustrate that this was applicable as a generic technique for increasing sensitivity (typically by around an order of magnitude) in low organic mobile phases.

Strategies to develop and validate methods for the determination of endogenous analytes in a biological fluid were investigated. This involved the use of a surrogate matrix, to develop a method for the determination of endogenous testosterone in human serum and the use of non-matrix calibration standards for the successful development and validation of a method for the analysis of indolyl 3 acryloylglycine (IAG) in human urine.

As a result of observations suggesting promotion of ionisation of deltamethrin in liver tissue sample extracts, it was postulated that this was due to the presence of high concentrations of surfactants. After confirming the effect, a series of systematic investigations were performed to attempt to understand the mechanism to be able to utilise this as a general method for the enhancement of signal with low sensitivity analytes. It was found that the type of surfactant and concentration used was directly associated with an increased (or decreased) response.

Although there remain a number of problems associated with the use of LC-API-MS, the work undertaken for this thesis has successfully demonstrated a number of techniques that can be applied to overcome these problems. Knowledge of the nature of the sample undergoing analysis, the required analytical conditions, and where required careful application of one of the techniques described will ensure that a robust method can be readily developed.

Item Type: Thesis (Doctoral)
Subjects: Sciences > Pharmacy and Pharmacology
Divisions: Collections > Theses
Depositing User: Barry Hall
Date Deposited: 10 Apr 2013 13:31
Last Modified: 20 May 2019 12:33

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